KR20180102868A - Composition comprising Vegetable Oil for preventing, alleviating or treatment of congnitive dysfunction disorder - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating, or treating cognitive dysfunction-related diseases containing color oil as an active ingredient. The color oil which is produced by using natural materials having high biocompatibility is aesthetically beautiful and improves cognitive ability efficiently, thereby being applied to various materials such as functional health foods and therapeutic agents for cognitive dysfunction-related diseases.

Description

[0001] The present invention relates to a composition for preventing, alleviating or treating cognitive dysfunction-related diseases,

The present invention relates to a composition for preventing, alleviating or treating a cognitive dysfunction-related disease comprising color oil as an active ingredient.

Fat is traditionally obtained from plants, animals, fish, etc. In particular, natural fat has various physical and chemical properties depending on the number of carbon atoms in the fatty acid, the degree of saturation of the fatty acid, the content of the isomer, and the polymorphic state of the fat. In general, edible oil is a generic term for commonly used oil (liquid at normal temperature) and fat (solid at normal temperature), and is derived from soybean oil, corn oil, vegetable oil, Sesame oil, sesame oil, safflower oil, sunflower oil, cottonseed oil, peanut oil, olive oil, palm oil, palm oil, red pepper seed oil and other edible oils.

Vegetable oil is generally called edible oil. Nutritionally, the calorie consumes 9 calories per gram, which is nearly double that of protein or carbohydrates. In Western countries, 35 to 40% of the energy intake is maintained through maintenance. Excessive intake of such fat causes health problems such as heart disease, obesity and cancer, and people's interest is increasing .

In recent years, interest in health and aesthetics of consumers has been increasing due to the economical margin of society. As a result, various functional additives are added to the conventional liquid edible oil to make various attempts according to consumers' Is continuing. At this time, it is important that the flavor inherent to the edible oil is not lowered by the additive.

The central nervous system, which consists of the brain and spinal cord, is a central center for the management of life phenomena. It is responsible for all functions of the human body, from sensory and (voluntary) exercises to thinking, memory, Is a very essential institution. Therefore, in all cases, such as the death of rapid nerve cells caused by stroke or trauma, or the death of slow nerve cells that cause degenerative diseases of the central nervous system such as senile dementia represented by Alzheimer's disease and Parkinson's disease, Resulting in an irreversible dysfunction of the network and eventually a permanent loss of the corresponding human function.

Elderly dementia, represented by Alzheimer 's disease, is characterized by an increase in the life expectancy of the human being and a proportional increase in association with the modernization of medical welfare facilities. According to the statistics of the Korea Institute for Health and Social Affairs, the elderly population of Korea exceeded 7% in 2000 and entered the aged society. In 2003, the number of elderly people reached 8.3%, 3.97% in 2003 and 14.4% . The prevalence of dementia in elderly people over 65 years of age is estimated to be 8.2%. In the western world, approximately 10% of the population aged 65 and over, 40-50% of the population aged 80 or older are developing Alzheimer's disease, and the US already has over 5 million people with this disease, It is estimated to be in dollars. In Korea, more than 200,000 people were found to be dementia patients. In the United States, it is estimated that it will double to 2030 by 2030 and reach more than 14 million by 2050 by more than 350%.

Accordingly, the present inventors have made intensive efforts to produce a color oil having a high content of natural pigments derived from natural materials and exhibiting a beautiful coloring property using the same, and having excellent functionality and functionality. As a result, the color oil of the present invention was completed. And has the purpose of preventing, alleviating or treating the cognitive dysfunction, and the present invention has been completed.

Korean Patent Laid-Open No. 10-2011-039762 Korean Patent Publication No. 10-2012-0016970

It is an object of the present invention to provide a composition for preventing, alleviating or treating cognitive dysfunction-related diseases comprising color oil as an active ingredient.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing, alleviating or treating cognitive dysfunction-related diseases comprising color oil as an active ingredient.

Further, the present invention provides a health functional food composition for preventing or alleviating cognitive dysfunction-related diseases comprising color oil as an active ingredient.

The color oil according to the present invention, which is manufactured using natural materials having high biocompatibility, is aesthetically beautiful and improves the cognitive ability efficiently, and can be utilized as various substances such as therapeutic agents for cognitive function disorders, health functional foods and the like .

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a view showing the appearance of a color oil of the present invention. FIG.
Fig. 2 is a graph showing the results of confirming the effect of improving the cognitive ability of color oil through the Y-maze experiment.
FIG. 3 is a diagram showing the results of confirming the effect of improving the memory ability and learning ability of color oil through the Morris water maze test.
FIG. 4 is a graph showing the results of confirming the activity of inhibiting the development of color oil and inhibiting lipid peroxidation through observation of changes in the content of acetylcholine esterase (AChE) and malondialdehyde (MDA).
FIG. 5 is a graph showing the results of confirming the antioxidative effect of color oil through changes in peroxide elimination enzyme (SOD) and glutathione (GSH) contents.

The present invention provides a composition for preventing, alleviating or treating cognitive dysfunction-related diseases comprising color oil as an active ingredient.

Hereinafter, the present invention will be described in more detail.

In the present invention, 'color oil' is a fat-oil produced by the present inventors, preferably 50 to 1500 μg of lycopene and 50 to 150 μg of beta-carotene per 100 g of color oil. The color oil exhibits a clear and beautiful colorless state without residue, has a high content of lycopene or benacarotin, has functionality such as antioxidant and antioxidant ability, and has an excellent effect.

In the present invention, the color oil is excellent in improving cognitive ability and memory effect, and thus can be effectively used for preventing or treating cognitive dysfunction.

The cognitive dysfunction-related diseases are selected from the group consisting of Alzheimer's disease, vascular dementia, Diffuse lewy body dementia, Huntington's disease, pick disease, Kruszfeld- Creutzfeldt-Jakob disease, dementia due to head injury, or Parkinson ' s disease.

In the present invention, the composition may contain, in addition to color oil, at least one known active ingredient having an effect of preventing or treating cognitive dysfunction.

The composition comprises a pharmaceutical composition and a food composition. Particularly, it includes health functional foods among food compositions.

In the present invention, the composition may further comprise suitable carriers, excipients and diluents conventionally used in the production of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral formulations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and the weight of the individual, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. For the desired effect, the rabbit extract of the present invention may be administered in an amount of 0.1 mg / kg to 50000 mg / kg per day, and may be administered once a day or divided into several times.

The pharmaceutical composition of the present invention can be administered to a subject in need thereof by various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers for the prevention and treatment of cognitive dysfunction.

In the present invention, the term "health functional food" refers to a food having a biological control function such as prevention and improvement of disease, bio-defense, immunity, recovery after disease and aging inhibition.

The composition of the present invention may be added to a health functional food for the purpose of preventing or improving cognitive dysfunction. When the rabbit extract of the present invention is used as a food additive, the ragweed extract can be used as it is or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the ragweed extract of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material, when the food or beverage is produced. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

There is no particular limitation on the kind of the food. Examples of the foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also provides, in yet another aspect, (a) a process for extracting an essential oil from a material containing lycopene or beta-carotene; And (b) mixing and stirring the essential oil obtained in step (a) with an edible oil, and stirring the oil.

In step (a), the material may be any of vegetables or fruits containing lycopene or beta-carotene. Examples of such materials include, but are not limited to, paprika, tomato, cherry, red pepper, Green pepper, green pepper, parsley, cucumber, lettuce, spinach, broccoli, and plum.

The refining substance extracted in the step (a) may be lycopene or beta-carotene.

In step (a), supercritical fluid extraction may be used to obtain lycopene or beta-carotene with high efficiency, and carbon dioxide is preferably used as the supercritical fluid. When supercritical fluid is extracted, carbon dioxide may be used alone, or an organic solvent may be mixed with an auxiliary solvent. The kind of the organic solvent is not limited thereto, but is preferably ethanol, more preferably at least 95% ethanol. If the concentration of the organic solvent is less than 95%, there is a problem that the water content of the extract is relatively high after concentrating the extract, thereby increasing the water content when added to the edible oil, and there is a fear that the stability of the product such as discoloration and precipitation is lowered . The mixing ratio of the carbon dioxide to the organic solvent may be 10: 1 to 2: 1.

The step (a) may be performed at a temperature of 30 to 60 ° C and a pressure of 100 to 300 bar, thereby preventing the destruction of the coloring component due to high-temperature extraction. The extraction time of the refinery material in step (a) is preferably 1 to 3 hours, more preferably 2 hours. If the extraction time is less than 1 hour, the extraction efficiency is significantly lowered. If the extraction time exceeds 3 hours, the extraction efficiency is not increased any more.

In the step (b), the edible oil is not limited to the group consisting of soybean oil, corn oil, rape seed oil, seed oil, sunflower oil, olive oil, palm oil, palm oil, safflower oil, sesame oil, perilla oil, peanut oil, It may be more than one selected. The edible oil is not browned, and the oiliness of the oil itself is not high, which is preferable for the production of the color oil of the present invention.

In the step (b), it is preferable that the refined material is contained in an amount of 1 to 5% by weight based on the total weight of the edible oil. If the content of the refining material is less than 1% by weight, the color saturation and specific odor are deteriorated. If the refining material content exceeds 5% by weight, the turbidity becomes high, and there is a fear of deposits.

Hereinafter, the present invention will be described in detail with reference to Examples and Production Examples. However, the following examples and preparations are merely illustrative of the present invention, and the content of the present invention is not limited by the following examples and production examples.

Example 1. Preparation of color oil

Lycopene and beta-carotene paprika, tomato, cherry, chili, omelet, watermelon, carrot, persimmon, orange, tangerine, kumquat, pumpkin, yellow jujube tomatoes, bell pepper, green pepper, cantaloupe, cucumber, lettuce, spinach, Broccoli, and plum were lyophilized and then ground to a particle size of about 60 mesh. The dried material was subjected to supercritical carbon dioxide treatment at 30 to 60 DEG C and 100 to 300 bar for 2 hours to extract an essential oil containing lycopene or beta carotene and 95% ethanol as an auxiliary solvent. The purified oil was mixed with edible oil and stirred to produce a finely colored oil shaking lycopene and beta carotene. The ratio of lycopene to beta carotene was adjusted to control the saturation of the color oil, so that red, orange, yellow, green Color oil was prepared.

More specifically, in order to prepare a red color oil, 5 g of an essential oil material extracted from at least one material selected from the group consisting of paprika, tomato, cherry, red pepper, omija and watermelon was added to 100 g of soybean oil as an oil for cooking.

Further, in order to prepare an orange color oil, 5 g of an essential oil material extracted from at least one material selected from the group consisting of carrot, persimmon and paprika was added to 100 g of soybean oil as an edible oil.

In addition, in order to prepare a yellow color oil, 5 g of an essential oil material extracted from at least one material selected from the group consisting of paprika, orange, mandarin orange, kumquat, amber and yellow jujubes was added to 100 g of edible oil.

In addition, 5 g of purified oil material extracted from at least one material selected from the group consisting of paprika, bell pepper, green pepper, parsley, cucumber, lettuce, spinach, broccoli and plum was added to 100 g of edible oil to prepare a green color oil.

The color of the color oil prepared through the above procedure was visually observed, and the results are shown in FIG.

Example  2. Through the Y-maze test color  Evaluating efficacy of improving cognitive ability of oil

2-1. Experimental animal

Four-week-old ICR-male mice were purchased from an experimental animal supplier (Samtako, Osan, Korea) and allowed to undergo a 7-day adaptation period. All animals were placed in one breeding case, , Relative humidity of 50-55%, and illumination time of 12 hours in the same laboratory environment. Experimental animals were divided into control group and trimethyltin chloride (TMT, Sigma-Aldrich Chemical Co., St. Louis, MO, USA) injection group (cognitive defect-induced control group), sample group (perilla oil and soybean oil Treated group) and divided into 6 groups of 9 mice per each group for 4 weeks. The sample group was orally administered daily with soybean oil and perilla oil (2.5 mL / kg body weight) for 21 days, and 100 μL of TMT (7.6 μg / kg body weight) dissolved in 0.85% saline on the 28th day To induce cognitive defects. After 2 days of TMT injection, the behavioral tests of Y-maze test and morris water maze test were conducted for 6 days.

2-2. Through the Y-maze test (Y-maze test) color  Identify the improvement of cognitive ability of oil

Y-maze experiment was performed 2 days after TMT injection. The Y-maze used in the experiment is made of black plastic and consists of three arms. The length, height and width of each arm are 33, 15, and 10 cm. After setting each arm to A, B, C, carefully place the mouse on one of the arms and move the arm's movement for 8 minutes using video-tracking system (Smart v3.0, Panlab SL, Barcelona, Spain) . 1 point (actual change) is assigned to each of three different arms in turn, and the alternation behavior is calculated using the following equation (1) using the total arm entry and score .

* Maximum alternation = total number of arm entry-2

 The Y-maze test is a method for evaluating the temporal spatial cognition of short-term memories. It uses a TMT that exhibits AD-like symptoms by causing irreversible damage to the hippocampus of the brain. The results of observing mouse behavior in Y-maze for 8 minutes are shown in Fig. The TMT group showed a decrease in memory (about 21%) compared to the control group (79%) and the soybean oil group (88%) showed no significant difference from the TMT group. In comparison with the TMT group, the perilla oil group was similar to the control group as well as improving memory. FIG. 3 shows the total number of times the mice passed through each arm of Y-maze for 8 minutes, and there was no significant difference in the basic athletic performance of the mice.

TMT-induced laboratory animals exhibit neurogenic hyperactivity resulting from limbic damage such as spontaneous epilepsy, aggression, and hypersensitivity. In Figure 2, the TMT group has significantly greater travel within the arm per hour compared to the control group Soybean oil group showed similar behavior. On the other hand, the perilla oil group showed no significant difference from the control group. Therefore, it shows the improvement effect of perilla oil on cognitive deficit and hyperactivity disorder induced by TMT abdominal injection.

Example  3. Through the Morris water maze test color  Confirming the improvement of memory and learning ability of oil

The Morris Underwater Labyrinth test can be used to measure spatial learning and cognitive enhancement, and is often used as a method to explore drugs for the prevention or treatment of dementia. The morris water maze experiment was performed the day after the Y-maze experiment of Example 2 to evaluate the spatial memory of the experimental animals. In a circular water tank (150 cm in diameter, 60 cm in height), the water was filled to a height of 30 cm (23 ± 2 ° C) and an escape platform was installed in a quadrant of the water tank and the edible inks (Cebesa, Valencia, Spain) . On the first day of the experiment, the experimental animals were allowed to swim freely for 60 seconds without a platform in the water tank, followed by adaptive training, and the next day, the platform remained visible by 1 cm above the water surface (visible trial). During the next 4 days, the platform was repeatedly trained once a day with different positions (N, S, E, W zone) obtained each time in a water tank set at 2 cm below the surface of the water. SL) (hidden trial). When the animal reached the platform in 60 seconds, it was allowed to stay on the platform for 10 seconds. If the animal could not be found, it was guided by the hand to position it on the platform for 20 seconds. On the fifth day of the experiment, a probe test was performed to record the time (in seconds) of staying in the zone (W zone) where the platform was located for 60 seconds to remove the platform and measure the working memory.

As a result, the Morris water maze experiment was performed to evaluate the spatial memory learning ability of the experimental animals. The results are shown in FIG. During the hidden trial, except for the soybean oil group, escape latency decreased with the passage of time. The TMT group had a longer time to visit the platform (W zone) than the control group, and the pericarp group had a longer time to visit than the control group on the first day, but decreased to a statistically significant level on the fourth day.

FIG. 3 shows a probe test for measuring spatial memory by measuring the time passed or staying in the zone (W zone) after platform was removed on the fifth day. The results are as follows. In the TMT group, 24.5% showed lower spatial memory than the control group (31.5%), and the soybean oil group (26.4%) showed spatial memory similar to the TMT group. On the other hand, the pericarp group (35.2%) had relatively good memory of the area where the platform was located, and the number of passages through the specific area where the platform was located was significantly larger ( P <0.05) showed that TMT improved spatial learning and memory impairment.

  Umezawa et al. Reported that sunflower seed oil and perilla oil, which are different in α-linolenic acid / linoleic acid (ω-3 / ω-6) fatty acid composition, were fed to mice with reduced learning and memory capacity (SAMP8) Perilla oil, which had a high ratio of α-linolenic acid, showed relatively good memory, learning and behavioral impairment. In animal studies using old mice, omega-3 fatty acid-deficient diets reduced spatial cognition and memory learning ability compared to the control group (14). Long-term ω-3 fatty acid-containing diets effectively improved memory impairment It has been reported that reducing the lipid peroxide content of the brain prevents the degenerative disorder caused by neuronal cell death (15). Soybean oil is composed of 85% of total fatty acids, unsaturated fatty acids, linoleic acid (65%), oleic acid (26%) and α-linolenic acid (9% (16%), linoleic acid (60%), oleic acid (17%) and linoleic acid (16). Therefore, the results of this experiment suggest that the relative difference in the content of ω-3 fatty acids partially influences the improvement of cognitive function.

Example  4. Acetylcholine Estherazade ( AChE ) Through confirmation of secretion amount color  Identification of neurotransmitter capacity of oil

After the completion of the Morris water maze test, 10 volumes of lysis buffer was added to the extracted mouse brain, homogenized with a glass-col homogenizer, centrifuged at 12,000 rpm for 30 minutes, and the supernatant was used for enzyme experiments. All extraction procedures were performed at 4 ° C and the protein content of the extracted enzyme was measured using the Quant-iT ™ protein assay kit (Invitrogen Co., Carlsbad, Calif., USA). After adding 65 μL of 50 mM sodium phosphate buffer to the enzyme, preincubate at 37 ° C for 15 minutes, add 70 μL of 250 mM substrate solution to the reaction mixture, incubate for 10 minutes, and insert into a microplate reader (Bio-Rad, Hercules, CA, USA Absorbance at 405 nm. AChE activity in mouse brain tissue was expressed as% activity relative to control group. The results are shown in Fig.

Enzymes involved in the synthesis and degradation of acetylcholine (ACh) are choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), respectively. Therefore, in order to facilitate neurotransmission of brain tissue, ChAT and AChE activity, which affect ACh levels, are very important. However, when AChE is activated as an ACh degrading enzyme, the concentration of ACh is decreased and the signal transmission of the brain is lowered, resulting in a decrease in cognitive ability. Therefore, a method of increasing the concentration of ACh by using AChE enzyme is being used in the treatment of degenerative dementia. One of the basic mechanisms of TMT-induced cranial nervous system damage is that TMT induces damage by inducing changes in cholinergic neuronal transmission through muscarinic receptor damage, which appears to occur in particular in the hippocampal neurons. As a result, TMT decreased the concentration of ACh in the hippocampal region. However, the direct correlation between TMT and AChE activity is not known, but it is considered to increase AChE activity in mouse brain tissue. Therefore, after the completion of the experiment of Y-maze and morris water maze, the brain of the mouse was extracted and the AChE activity was measured. TMT group showed AChE activity activity increased to 148% compared to control group (100%) and 103% in perilla oil group, showing 30% inhibition effect compared with TMT group. The soybean oil group inhibited AChE activity to about 140%, but the effect was insignificant when compared with perilla oil.

Example  5. Malondialdehyde ( MDA ) Through content change color  Identification of oil lipid peroxidation inhibitory activity

 After adding phosphate buffered saline to the extracted mouse brain, 160 μL of the obtained homogenate was mixed with 960 μL of 1% phosphoric acid. Then, 320 μL of 0.67% thiobarbituric acid was added and reacted at 95 ° C. for 1 hour. The reaction solution was centrifuged at 5,000 rpm for 10 minutes in a centrifuge (Combi-514R, Hanil Co. Ltd., Seoul, Korea), and the absorbance of the supernatant (Libra S32PC, Biochrom Ltd., Cambridge, UK) was measured at 532 nm . The MDA content was expressed as the concentration of nmole per mg protein. The results are shown in Fig.

The peroxidation phenomenon in the cells constituting the tissue is caused by binding to the free radicals of the unsaturated fatty acids constituting the main constituent phospholipid of the cell membrane and the lipid peroxidation is a by-product in the unsaturated fatty acids and lipoproteins of the cell membranes, resulting in a malondialdehyde (MDA ). MDA is known to be increased in various regions of brain tissue during oxidative stress and has been reported to cause oxidative damage to tissues such as DNA and condensation during accumulation. In particular, brain cells are more susceptible to oxidative damage due to higher content of unsaturated fatty acids compared to other organs. When oxidative stress is applied to brain nerve cells, they cause neurotoxicity after inflammatory reaction and promote nerve cell death. In the end, this oxidative stress is expected to cause memory impairment and loss due to persistence in brain tissue. In this experiment, MDA content of each group was measured in order to compare the content of MDA, a lipid peroxide intermediate present in brain tissue of mice fed soybean oil and perilla oil. The control group was 4.21 nmole / mg protein, and the TMT group showed an increase of 5.45 nmole / mg protein. MDA content of soybean oil and perilla oil groups were 4.64 and 3.20 nmole / mg protein, respectively, which inhibited MDA production compared to TMT group, but soybean oil did not show significant difference, while perilla oil had about 41% inhibitory effect And MDA content was significantly lower than control. The perilla oil contains physiologically active substances such as luteolin, apigenin, caffeic acid, ascorbic acid, β-carotene, limonene and protocatechuic acid, which are known to have very high anticancer properties. In Korea, it is reported that perilla oil is used without refining differently from common cooking oil. Therefore, it is reported that such physiologically active substances are relatively abundant and that lipid oxidation in mouse brain tissue can be inhibited by the antioxidant effect thereof. Therefore, TMT seems to damage neurons in specific areas of the brain, suggesting that TMT - induced damage to brain nerve cells is more effective than that of soybean oil.

Example  6. Peroxide Removal Enzyme ( superoxide dismutase ; SOD) activity of color oil

The brain of the extracted mouse is homogenized with 10 volumes of lysis buffer, and homogenized with a glass-col homogenizer. Centrifuge at 12,000 rpm for 30 minutes to discard the supernatant and take a pellet. 1X Cell Extraction Buffer (1 mL of 10X SOD Buffer, 0.2 mL of 20% triton X-100, 8.8 mL of distilled water, 10 μL of 200 mM pMSF) was vortexed for 30 minutes at 5 minutes and then centrifuged at 1,000 rpm for 10 minutes The supernatant was used for the experiment. All procedures were maintained at 4 ° C and the Quant-iT ™ protein assay kit (Invitrogen Co) was used to determine the protein content of the supernatant. SOD activity was measured by SOD determination kit (Sigma-Aldrich Chemical Co.). Absorbance was calculated and expressed as SOD (U / mL). The results are shown in Fig.

One of the antioxidant enzymes, SOD, is an enzyme that catalyzes the conversion of reductive oxygen species that are harmful to cells into hydrogen peroxide. H ₂ O ₂ produced by SOD is converted into harmless water molecules and oxygen molecules by peroxidase or catalase It is known to protect the body from oxygen injury. In addition, SOD is an enzyme that essentially eliminates free radicals, and has been shown to be more effective than other antioxidants, resulting in a great deal of interest as a pharmaceutical preparation. Currently, SOD is used in cosmetics as an anti-inflammatory agent to be. The SOD assay kit-WST used in this study was composed of highly water-soluble tetrazolium salt and WST-1 (2- (4-iodophenyl) -3- (4- nitrophenyl) -5- (2, -disulfi-phenyl) -2H-tetrazolium, monosodium salt. WST-1 reacts with superoxide anion to produce water-soluble formazan dye, while WST-1 does not react with the reduced form of xanthine oxidase, and its reactivity with superoxide anion is 70 times lower than that of cytochrome C, High sensitivity is shown even in low concentration samples. The control group was 16.5 U / mg protein and the TMT group was 11.7 U / mg protein. The SOD contents of soybean oil and perilla oil groups were 12.6 and 14.2 U / mg protein, respectively. The soybean oil showed little difference from the TMT group, but the perilla oil group effectively inhibited the decrease of SOD.

Example  6. Glutathione ( glutathine ; GSH ) Through content change color  Oil Oxidative  Confirmation of cytotoxic effect

After the homogenization of 10 volumes of cold 5% metaphosphoric acid was added to a certain amount of mouse brain, the supernatant was obtained by centrifugation (14,000 × g) for 15 minutes and 10 μL of 2 M 4-vinylpyridine was added to measure oxidized glutathione Respectively. The whole process was maintained at 4 ° C and Quant-iT ™ protein assay kit (Invitrogen Co) was used to measure the protein content of the extracted supernatant. Glutathione content was measured using a glutathione (GSSG / GSH) detection kit (Enzo Diagnostics, Farmingdale, NY, USA) and the absorbance values of the measured total glutathione and oxidized glutathione (GSH) and total GSH (%). The results are shown in Fig.

Glutathione in brain tissue is a substance used as a substrate of GSH-peroxidase and GST, which are enzymes that protect against oxidative cell damage. It is a process in which active oxygen, lipid peroxides and electrophilic substances are finally detoxified in cells And carries out various functions such as elimination of lipid peroxidation substances and foreign substances in cells, amino acid transport and storage, and liver decryption. In the control group, the amount of glutathione oxidized in total glutathione was 8.4%, that of TMT group was 10.4%, that of oxidized glutathione increased in the control group, 5.3% in the perilla oil group, Showed excellent antioxidant effect of glutathione. On the other hand, the ratio of oxidized glutathione was higher than that of TMT group in soybean oil group (14.6%). Vitamin A, E, and C were fed to rats exposed to oxidative stress. Vitamin E showed the highest levels of SOD and GSH in brain tissues. In conclusion, It is thought that this is due to the abundance of physiologically active components including vitamin E relative to other oils including edible oil.

The results were expressed as means and standard deviations (mean ± SD), and the statistical significance of differences between the experimental groups was analyzed using ANOVA variance analysis and Duncan's multiple range test (SAS Institute Inc., Cary, NC, USA) ( P <0.05). There was no significant difference between the two groups.

Hereinafter, a preparation example of a composition for preventing, alleviating or treating cognitive dysfunction comprising the color oil of the present invention as an active ingredient will be described, but the present invention is not intended to be limited thereto but is specifically described.

Formulation Example 1. Preparation of pharmaceutical preparations

1. Manufacturing of powder

Color oil 20 mg

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

2. Preparation of tablets

Color oil 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

3. Preparation of capsules

Color oil 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

4. Preparation of injections

Color oil 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

5. Manufacture of liquids

Color oil 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation Example 2. Preparation of food preparation

1. Manufacture of health food

Color oil 100 mg

Vitamin mixture quantity

70 g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 g of vitamin B12

Vitamin C 10 mg

Biotin 10 g

Nicotinic acid amide 1.7 mg

Folate 50 g

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2. Manufacture of health drinks

Color oil 100 mg

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the health beverage composition of the invention.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (2)

A pharmaceutical composition for preventing, alleviating or treating cognitive dysfunction-related diseases comprising color oil as an active ingredient. A health functional food composition for preventing or alleviating cognitive dysfunction related diseases comprising color oil as an active ingredient.

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